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Volume 10,Issue 4

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26 December 2025

Validation Strategy for Monoclonal Antibody Purity Detection Methods Based on Risk Assessment - a Review of the Core Technical Requirements of the 2025 Version of ChP and USP

Xiaoxiao Fu1,3 Huaiyao Qiao2 Fang Wang3 Jie Gu3 Ana Chen1*
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1 Anhui Engineering University, Wuhu 241000, Anhui, China
2 Jiangsu Quanxin Biopharmaceutical Co., Ltd., Taizhou 225300, Jiangsu, China
3 Taizhou, Jiangsu 225300; Jiangsu Saifus Biotechnology Co., Ltd., Taizhou 225300, Jiangsu, China
JMDS 2025 , 10(4), 193–199; https://doi.org/10.18063/JMDS.v10i4.1207
© 2025 by the Author. Licensee Whioce Publishing, Singapore. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution -Noncommercial 4.0 International License (CC BY-NC 4.0) ( https://creativecommons.org/licenses/by-nc/4.0/ )
Abstract

Verification of monoclonal antibody (mAb) purity detection methods is a key link to ensure drug quality. However, in the current validation practice of size exclusion high-performance liquid chromatography (SEC-HPLC) and capillary electrophoresis (CE), there are outstanding problems such as inconsistent technical implementation and "formalization" of projects, and there are many disputes over the selection of validation projects. Among them, the SEC-HPLC area normalization method is the most widely used semi-quantitative analysis method for polymers in enterprises. The selection and implementation of verification projects are particularly chaotic. At the same time, methods such as ion exchange chromatography (IEC) often fail to achieve baseline separation, and their verification logic has also become the focus of the industry. This article is based on the 2025 version of the Pharmacopoeia of the People's Republic of China (ChP 2025), the latest specifications of the United States Pharmacopoeia (USP) and the results of special literature research, systematically sorting out the chaos and core disputes in enterprise verification, focusing on interpreting the verification project design logic of the SEC-HPLC area normalization method, explaining the core reasons for choosing this method for purity testing of biological products, and focusing on IEC that does not reach baseline separation. The testing method proposes verification ideas, starting from the perspective of method characteristics, pharmacopoeia requirements, and risk control, and summarizes a unified verification plan design idea to provide enterprises with a technical reference that is both compliance and practical, and promote the standardized development of industry verification practices.

Keywords
monoclonal antibody
purity detection
method validation
SEC-HPLC
area normalization method
ion exchange chromatography
References

[1] National Pharmacopoeia Committee, 2025, Pharmacopoeia of the People’s Republic of China (Part Four) (2025 edition). China Medical Science and Technology Press.

[2] United States Pharmacopeial Convention, 2025, USP <129> Analytical Procedures for Recombinant Therapeutic Monoclonal Antibodies. USP Convention.

[3] United States Pharmacopeial Convention, 2025, USP <621> Chromatography. USP Convention.

[4] United States Pharmacopeial Convention, 2025, USP <1053> Capillary Electrophoresis for Biotechnology Products. USP Convention.

[5] United States Pharmacopeial Convention, 2025, USP <1225> Validation of Compendial Procedures. USP Convention.

[6] Guo W, Ding L, Zhang M, et al., 2014, Comparative Study on the Chromatographic Behavior of TNF-α Monoclonal Antibody Molecular Size Variants and Charge Variants. Journal of Pharmaceutical Analysis, 34(8): 1425-1430.

[7] Xu GL, Li M, Wang C, et al., 2024, Validation Study of Monoclonal Antibody Reduction Capillary Electrophoresis Method. Chinese Journal of New Drugs, 33(5): 489-495.

[8] International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use, 2022, ICH Q2(R1) Validation of Analytical Procedures: Text and Methodology.

[9] National Medical Products Administration, 2024, 2025 Drug Testing Proficiency Testing Plan. visited on November 20, 2025. https://www.nmpa.gov.cn/xxgk/fgwj/gzwj/202412/t20241210_176542.html.

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